Structure-function relationship in the antifreeze activity of synthetic alanine-lysine antifreeze polypeptides.

Recently antifreeze proteins (AFP) have been the subject of many structure−function relationship studies regarding their antifreeze activity. Attempts have been made to elucidate the structure−function relationship by various amino acid substitutions, but to our knowledge there has been no successful from first principles design of a polypeptide that would bind to designated ice planes along a specific direction. In this paper we show the results of our first attempt on an entirely de novo design of an alanine−lysine-rich antifreeze polypeptide. This 43 residue alanine−lysine peptide exhibits characteristic nonequilibrium freezing point depression and binds to the designated (210) planes of ice along the [122] vector. The structural and thermodynamic properties of this polypeptide were determined using circular dichroism spectroscopy and its nonequilibrium antifreeze properties were investigated using an ice-etching method and nanoliter osmometry.